Protocols

A practical discussion on creating yeast lysate

Yeast cell lysis protocol, glass beads and vortexer
Definition of grams of wet weight cells
Estimating percentage cell disruption

Cellcrusher has developed the Spiral Mill, a unique cooled bead beater, which offers a faster, easier, temperature controlled alternative to the classic method of disrupting cells by agitating with glass beads on a vortexer.  Our device is specifically designed for mid-size (1 g to 6 g wet weight) samples.

However, we understand that researchers may wish to conduct preliminary work on small yeast and fungus samples, before they commit resources to buying a specialised cell-breaking machine for larger scale, temperature controlled, reproducible work.  Therefore, we have provided the following yeast cell lysis protocol, using glass beads, a vortexer and centrifuge tubes.

These protocols and discussions are written in an informal style, in order to better explain the rationale behind the approaches recommended.

Yeast cell lysis protocol, glass beads and vortexer

This protocol provides detailed instructions on breaking 1 g wet weight of yeast in a 15 ml centrifuge tube.  Also included is a discussion on potential problems associated with disruption of yeast, algae and fungus using this technique.

Definition of grams of wet weight cells

This discussion focuses on the importance of how the wet weight of cell pellets are defined.  Difficulties in definition stem from the fact that cell pellets shrink significantly after freezing and thawing.  Inaccurate estimates size of pellets can lead to poor disruption.

Estimating percentage cell disruption

When optimizing extraction protocols for yeast/fungus in unicellular form, an essential first step is to use a hemacytometer and microscope to do a quick estimation of cell lysis.  This protocol and accompanying photograph of broken yeast cells (also known as “ghosts”) is intended to assist in this task.